Animals ; Antibodies, Monoclonal ; metabolism ; therapeutic use ; Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; therapy ; Drug Delivery Systems ; Epidermal Growth Factor ; immunology ; metabolism ; physiology ; Epithelial-Mesenchymal Transition ; GPI-Linked Proteins ; Humans ; Immunotherapy ; methods ; Intercellular Signaling Peptides and Proteins ; Membrane Glycoproteins ; immunology ; metabolism ; physiology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Proteins ; immunology ; metabolism ; physiology

Objective To understand current situation of surgery grading management in 22 hospitals from Beijing Municipal Administration of Hospitals,and to discuss the problems and to make suggestions for better application and management.Methods Based on general survey,Surgery of Grading Conditions of All Municipal Hospitals was issued to 22 municipal hospitals to make related staff fill related data.Results Except for 3 specialty hospitals,19 hospitals have launched the surgery grading management,while they are in different steps of implementing hierarchical directory,surgeon permission and dynamic management.These hospitals have different reference standards of making hierarchical directory.84.2％ hospitals give surgeons authorities according to levels of operations evaluated by professional title,surgery grade and doctors' technical skills.There are 14 hospitals applying information management on surgery grading.Conclusion Uniform criteria of hierarchical directory,extension of surgeon permission system and implementation of surgery grading information management will contribute to regulation fulfillment.

Objective To diagnose the first imported case of Plasmodium ovale infection by laboratory detection. Meth?ods The epidemiological data and blood samples of the case were collected,and the samples were detected by the microscopic examination,rapid diagnostic test(RDT)and nested PCR. Results The patient was a construction worker backing from Con?go,Africa. He experienced the symptoms of irregular fever and weakness one month after returning in Lingyang Town,Junxian County. The results of RDT only suggested no?Plasmodium falciparum infection. Under the microscope,it was seen that the in?fected RBC were obviously disfigured and in irregular shape,the ring forms were thick and big,and also thick granulas in big trophozoite stage and schizont stage were found. The results of PCR showed that the size of amplified product was about 800 bp, which was conformed to that of P. ovale. Conclusion Though microscopic examination is the golden standard for malaria diag?nosis,as P. ovale is difficult to be identified under microscope,the microscopic method combined with PCR test can be used for definite diagnosis.

Objective To investigate the effects of cyclooxygenase-2 inhibitor celecoxib on proliferation , invasion and migration of human pancreatic cancer cell line PANC-1 and then determine the optimal concentration of cele-coxib and the most suitable application time .Methods Human pancreatic cancer cell line PANC-1 was treated with diverse concentrations of celecoxib (20,60,100 μmol/L) for different durations (24,48,72 h).Cell prolifer-ation, invasion and migration capabilities were measured by MTT colorimetry , Transwell invasion assay , and scratch assay respectively .Results The proliferation capability of PANC-1 cell was reduced by celecoxib in a con-centration-and time-dependent manner ( P <0.05 ) .In addition , the invasion and migration capabilities were decreased by celecoxib in a concentration-dependent manner(P<0.01).Conclusions Celecoxib attenuates the proliferation of human pancreatic cancer cell line PANC-1 in a concentration-and time-dependent manner .Cele-coxib attenuates the invasion and migration in a concentration-dependent manner .

Objective To investigate the anti-tumor effect and mechanism of curcumin in pancreatic cancer (PC). Methods Smad4 and Jab1 expressions were detected by immunohistochemistry in tumor tissues and pericarcinomatous tis?sue from 35 PC cases, and the correlation of Smad4 and Jab1 were analyzed based on the percentage of positive staining in?tissues from 21 random selected PC cases. The effect of curcumin on expressions of tumor suppressors p53, Smad4 and cell cycle inhibitor p27 were examined by Western Blotting after human pancreatic cancer cell line PANC-1 were divided into PANC-1 control group (no treatments were given) and PANC-1 curcumin group (treated with cell culture medium containing 10μmol/L curcumin). The effect of curcumin on expressions of combination of β-TrCP1 and Smad4 was examined by Co-Immunoprecipitation after human embryonic kidney cell line 293T were divided into 293T control group (no treatments were given), 293T curcumin group (treated with cell culture medium containing 10μmol/L curcumin) and 293T Jab1 group (trans?fected by HA-Jab1 plasmid). Results Compared with expressions in pericarcinomatous tissues, Smad4 was down regulated while the expression of Jab1 was upregulated in PC tissues (P<0.01), and the expression of Smad4 was negatively correlated with the expression of Jab1 (n=21, r=-0.71, P=0.007). After treated with curcumin, the protein expression of p53, Smad4 and p27 was increased in PANC1 cell, and the protein expression of the combination ofβ-TrCP1 and Smad4 was decreased in 293T cell (P<0.05). After transfected by HA-Jab1 plasmid, the protein expression of the combination ofβ-TrCP1 and Smad4 was increased in 293T cell (P<0.05). Conclusion Curcumin may have suppression effect of PC through increasing the protein expression of p53, Smad4 and p27, and the mechanism of Smad4 upregulation may be related with the inhibition of Smad4 ubiquitination process, while Jab1 may be also involved in Smad4 degradation through ubiquitination.

Objective To explore the expression of matrix metalloproteinase-14(MMP-14)protein in the human stomach carcinoma tissues and its correlation with carcinoma invasiveness and metastasis.Methods The MMP-14 protein expression was detected by immunohistochemistry in 59 cases of stomach carcinoma tissues (observation group)and 20 cases of normal stomach tissues (control group,the adjacent normal tissues from the tumor margin of 5 cm confirmed by pathology),and its correlation with the clinically pathological parameters was analyzed.The expression characteristics of MMP-14 among various TNM stages of stom-ach carcinoma were also analyzed.Results The positive rate of MMP-14 expression was 50.85%(30/59)in the observation group and 5.00% (1/20)in the control group,the positive rate of the observation group was significantly higher than that of the control group (P <0.01);the expression level of MMP-14 was correlated with the differentiation degree,regional lymph node metastasis degree,invasion depth,lymphatic invasion and TNM stage,which showing the statistical difference(P < 0.01);the expression of MMP-14 protein was up-regulated and showed the transferring trend from cytoplasm to cellular membrane along with the progres-sion of TNM stage.Conclusion The overexpression of MMP-14 protein exists in stomach carcinoma tissues,which contributing to the invasion and metastasis of stomach carcinoma cells.

The HPLC fingerprint determination method of Jinzhen oral solution was established to provide a new method for quality control of Jinzhen oral solution. RP-HPLC was used for phenomenex Luna C18 (4.6 mm x 250 mm, 5 μm) chromatographic column, with 0.1% H3 PO4 water solution and acetonitrile as the mobile phase for gradient elution. The detection wavelength was 280 nm. HPLC fingerprint of Jinzhen oral solution was established to identify 17 common peaks in Jinzhen oral solution. The similarity of fingerprints of 10 batches of finished products was more than 0. 90. The established HPLC fingerprint has a better precision, reproducibility and stability, and can be applied in quality control of Jinzhen oral solution.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control

Signal transducer and activator of transcription 3 (STAT3) is an important regulatory factor of cell proliferation and metastasis, involved in the occurrence and development of a variety of malignant tumors, and it is one of the hot spots in the research of targeted anti-tumor drugs. Our group screened a novel benzobis (imidazole) structure small molecule compound LZJ541 through the screening model of Janus kinase (JAK)/STAT3 pathway inhibitors, which has definite STAT3 inhibitory activity. We examined the effect of LZJ541 on the proliferation of HepG2 and PC-3 cells by MTT assay in vitro, detected the effect of LZJ541 on the expression of STAT3-related proteins in HepG2 cells by Western blot, and measured the effect of LZJ541 on the apoptosis and cell cycle arrest of HepG2 cells via flow cytometry. The results indicated that LZJ541 significantly inhibited the activation of STAT3 signaling pathway and restrained the proliferation of HepG2 cells. Its half maximal inhibitory concentration (IC₅₀) was 13.8 μmol·L^-1, which was much lower than that of PC-3 cells (with low STAT3 expression, IC₅₀: 41.99 μmol·L^-1), LZJ541 can also inhibit the phosphorylation of STAT3 in HepG2 cells, thereby inducing apoptosis and cycle arrest and then exerting anti-tumor effects. In conclusion, LZJ541 has a certain anti-tumor effect in vitro, which provides an experimental basis for the development of new STAT3-targeted anti-tumor drugs around this kind of compounds.

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

OBJECTIVE To investigate the expression and significance of Fas, FasL and FADD in laryngeal carcinoma. METHODS Immunohistochemical method was used to examine the expression of Fas, FasL and FADD in laryngeal carcinoma specimen and adjacent normal tissues. RESULTS The positive rates of Fas and FADD in laryngeal carcinoma tissue were significantly lower than that in adjacent normal tissue, while FasL in laryngeal carcinoma tissue was higher than that in adjacent normal tissue(P